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Ubiquitination of plasma membrane ectophosphatase in bloodstream forms of Trypanosoma brucei

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Abstract

Bloodstream forms of Trypanosoma brucei contain plasma-membrane-integral acidic ectophosphatase. Here, it is shown by N-terminal sequencing that the ectophosphatase found in ricin-binding material was modified by ubiquitin. Three different ubiquitinated species corresponding to single, double and triple ubiquitinated forms of the enzyme were identified. Immunofluorescence studies with live bloodstream-form parasites showed that the ectophosphatase was localized in the flagellar pocket—the sole site for endocytosis in trypanosomes. As ubiquitin modification of plasma membrane proteins serves as an internalization signal, it is suggested that ubiquitinated ectophosphatase is labelled for endocytosis.

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References

  • Bakalara N, Santarelli X, Davis C, Baltz T (2000) Purification, cloning, and characterization of an acidic ectoprotein phosphatase differentially expressed in the infectious bloodstream form of Trypanosoma brucei. J Biol Chem 275:8863–8871

    Article  PubMed  CAS  Google Scholar 

  • Baltz T, Baltz D, Giroud C, Crockett J (1985) Cultivation in a semi-defined medium of animal infective forms of Trypanosoma brucei, T. equiperdum, T. evansi, T. rhodesiense and T. gambiense. EMBO J 4:1273–1277

    PubMed  CAS  Google Scholar 

  • Brickman MJ, Balber AE (1993) Trypanosoma brucei rhodesiense: membrane glycoproteins localized primarily in endosomes and lysosomes of bloodstream forms. Exp Parasitol 76:329–344

    Article  PubMed  CAS  Google Scholar 

  • Cross GAM (1975) Identification, purification and properties of clone-specific glycoprotein antigens constituting the surface coat of Trypanosoma brucei. Parasitology 71:393–417

    Article  PubMed  CAS  Google Scholar 

  • Dupré S, Urban-Grimal D, Haguenauer-Tsapis R (2004) Ubiquitin and endocytic internalization in yeast and animal cells. Biochim Biophys Acta 1695:89–111

    Article  PubMed  CAS  Google Scholar 

  • Haglund K, Di Fiore PP, Dikic I (2003) Distinct monoubiquitin signals in receptor endocytosis. Trends Biochem Sci 28:598–603

    Article  PubMed  CAS  Google Scholar 

  • Hicke L, Dunn R (2003) Regulation of membrane protein transport by ubiquitin and ubiquitin-binding proteins. Annu Rev Cell Dev Biol 19:141–172

    Article  PubMed  CAS  Google Scholar 

  • Hirumi H, Hirumi K, Doyle JJ, Cross GAM (1980) In vitro cloning of animal-infective bloodstream forms of Trypanosoma brucei. Parasitology 80:371–382

    PubMed  CAS  Google Scholar 

  • Horák J (2003) The role of ubiquitin in down-regulation and intracellular sorting of membrane proteins: insight from yeast. Biochim Biophys Acta 1614:139–155

    Article  PubMed  CAS  Google Scholar 

  • Lanham SM, Godfrey DG (1970) Isolation of salivarian trypanosomes from man and other animals using DEAE-cellulose. Exp Parasitol 28:521–534

    Article  PubMed  CAS  Google Scholar 

  • Li Z, Zou C-B, Yao Y, Hoyt MA, McDonough S, Mackey ZB, Coffino P, Wang CC (2002) An easily dissociated 26 S proteasome catalyzes an essential ubiquitin-mediated protein degradation pathway in Trypanosoma brucei. J Biol Chem 227:15486–15498

    Article  CAS  Google Scholar 

  • Lowrie DJ Jr, Giffin BF, Ventullo RM (1993) The ubiquitin–ligase system in Trypanosoma brucei brucei. Am J Trop Med Hyg 49:545–551

    PubMed  CAS  Google Scholar 

  • Overath P, Chaudhri M, Steverding D, Ziegelbauer K (1994) Invariant surface proteins in bloodstream forms of Trypanosoma brucei. Parasitol Today 10:53–58

    Article  PubMed  CAS  Google Scholar 

  • Overath P, Stierhof Y-D, Wiese M (1997) Endocytosis and secretion in trypanosomatid parasites—tumultuous traffic in a pocket. Trends Cell Biol 7:27–33

    Article  PubMed  CAS  Google Scholar 

  • Pickart CM (2001) Mechanisms underlying ubiquitination. Annu Rev Biochem 70:503–533

    Article  PubMed  CAS  Google Scholar 

  • Rodriguez J, Deinhardt F (1960) Preparation of a semipermanent mounting medium for fluorescent antibody studies. Virology 12:316–317

    Article  PubMed  CAS  Google Scholar 

  • Rotin D, Staub O, Haguenauer-Tsapis R (2000) Ubiquitination and endocytosis of plasma membrane proteins: role of Nedd4/Rsp5p family of ubiquitin–protein ligases. J Membr Biol 176:1–17

    Article  PubMed  CAS  Google Scholar 

  • Staub O, Gautschi I, Ishikawa T, Breitschopf K, Ciechanover A, Schild L, Rotin D (1997) Regulation of stability and function of the epithelial Na+ channel (ENaC) by ubiquitination. EMBO J 16:6325–6336

    Article  PubMed  CAS  Google Scholar 

  • Steverding D, Stierhof Y-D, Chaudhri M, Ligtenberg M, Schell D, Beck-Sickinger AG, Overath P (1994) ESAG 6 and 7 products of Trypanosoma brucei form a transferrin binding protein complex. Eur J Cell Biol 64:78–87

    PubMed  CAS  Google Scholar 

  • Strous GJ, Govers R (1999) The ubiquitin–proteasome system and endocytosis. J Cell Sci 112:1417–1423

    PubMed  CAS  Google Scholar 

  • Télles S, Abate T, Slezynger TC, Henríquez DA (1999) Trypanosoma cruzi and human ubiquitin are immunologically distinct proteins despite only three amino acid difference in their primary sequence. FEMS Immunol Med Microbiol 24:123–130

    Article  PubMed  Google Scholar 

  • Van Meirvenne N, Janssens PG, Magnus E (1975) Antigenic variation in syringe passaged populations of Trypanosoma (Trypanozoon) brucei. I. Rationalization of the experimental approach. Ann Soc Belge Med Trop 55:1–23

    Google Scholar 

  • Webster P, Russell DG (1993) The flagellar pocket of trypanosomatids. Parasitol Today 9:201–206

    Article  PubMed  CAS  Google Scholar 

  • Ziegelbauer K, Multhaup G, Overath P (1992) Molecular characterization of two invariant surface glycoproteins specific for the bloodstream stage of Trypanosoma brucei. J Biol Chem 267:10797–10803

    PubMed  CAS  Google Scholar 

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Acknowledgement

I thank Bianca Hube, Katrin Kremp and Nicole Tilly for excellent technical assistance, and Dr. Darren Sexton for critical reading of the manuscript.

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Correspondence to D. Steverding.

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This work was supported by the Bundesministerium für Bildung, Wissenschaft, Forschung und Technik, Schwerpunkt für tropenmedizinische Forschung in Heidelberg (01 KA 9301/3).

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Steverding, D. Ubiquitination of plasma membrane ectophosphatase in bloodstream forms of Trypanosoma brucei . Parasitol Res 98, 157–161 (2006). https://doi.org/10.1007/s00436-005-0045-3

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